SRM Assay Day 9

My samples are DONE.

The samples I needed to rerun to get data for (O01, O12, O22, O113, O118, O26 and O90) just finished running and all of them have data. Tomorrow, I will go to UWPR to prepare the oyster sample dilutions needed to create a dilution curve. The purpose of this is to verify that the assay is detecting peptides based on their presence in the autosampler vial. The oyster peptide detection should decrease as the concentration in the autosampler vial decreases, and vice versa.

Emma suggested pooling oyster samples to create my dilutions. I want to randomly pick oyster samples that I didn’t have to remake and rerun. I also don’t want to include my procedural blanks. Here are the samples I can choose from:

O17

O145

O128

O56

O71

O103

O04

O106

O49

O52

O14

O102

O10

O06

O122

O99

O08

O51

O21

O39

O137

O46

O43

O31

O66

O30

O100

O78

O35

O101

O40

O64

O91

O140

O124

O32

O147

O121

O24

O96

O131

O60

We want ten dilutions total. I first need to calculate how much of the pooled oyster protein sample I’ll need in each vial, as well as PRTC.

Table 1. Volumes of solutions needed to create peptide dilutions.

Vial

Oyster Sample Volume (µL)

Volume PRTC (µL)

Remaining Volume (µL)

1

7.5

1.89

5.61

2

6.7

1.89

6.41

3

5.9

1.89

7.21

4

5.1

1.89

8.01

5

4.3

1.89

8.81

6

3.5

1.89

9.61

7

2.7

1.89

10.41

8

1.9

1.89

11.21

9

1.1

1.89

12.01

10

0

1.89

13.11

This means I need 38.7 µL of oyster peptide sample to start with. If I used five samples, I would need 8 µL of peptide from each sample.

Randomly choosing five samples:

O124

O39

O96

O14

O137

I will prepare my samples accordingly tomorrow! Before then, I also did some “maintenance” for my Skyline stuff: